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[目的]体外观察TZDs类药物对小鼠原代骨髓细胞向成骨细胞和破骨细胞分化的影响,探讨其对糖尿病患者骨骼代谢的影响和作用机制。[方法]无菌条件下从小鼠股骨分离获取骨髓细胞,贴壁细胞进行成骨细胞生成实验研究,悬浮细胞进行破骨细胞生成实验研究。随后在1、2、5μmol/L罗格列酮干预下诱导成骨细胞分化培养3周,进行Von Kossa染色观察成骨细胞矿化结节,并测定成骨细胞标记物ALP活性、BMP-2及TGF-β分泌的影响;观察罗格列酮对破骨细胞形成和成熟的影响及标志性基因表达。[结果]1、2、5μmol/L罗格列酮干预组与对照组相比,成骨细胞的钙结节形成比例显著降低(P<0.05),ALP、BMP-2、TGF-β水平呈剂量依赖性地下降(均P<0.05);同时剂量依赖性促进c-fos和RANK基因的表达,破骨细胞生成实验显示罗格列酮促进破骨细胞分化发育和成熟。[结论]罗格列酮可剂量依赖性地抑制骨髓细胞向成骨细胞分化,促进向破骨细胞分化。有助于理解Ⅱ型糖尿病患者在应用TZDs骨丢失的原因。
[Objective] To observe the effect of TZDs on differentiation of mouse primary bone marrow cells into osteoblasts and osteoclasts in vitro and to explore its effect on bone metabolism in diabetic patients and its mechanism. [Method] The bone marrow cells and adherent cells were isolated from mouse femurs under aseptic conditions for osteoblastogenesis experiments and osteoclastogenesis experiments on suspension cells. Subsequently, osteoblasts were induced to differentiate and cultured for 3 weeks under the influence of 1, 2, 5μmol / L rosiglitazone. Von Kossa staining was used to observe the mineralized nodules of osteoblasts. ALP activity, osteoblast marker BMP-2 And TGF-β secretion; observed Rosiglitazone on the formation and maturation of osteoclasts and gene expression. [Results] Compared with the control group, the proportion of calcium formation in osteoblasts in 1,2,5 μmol / L rosiglitazone group was significantly decreased (P <0.05), while the levels of ALP, BMP-2 and TGF-β (P <0.05). At the same time, the expression of c-fos and RANK genes was promoted in a dose-dependent manner. The osteoclastogenesis experiments showed that rosiglitazone promoted osteoclast differentiation and maturation. [Conclusion] Rosiglitazone inhibits the differentiation of bone marrow cells into osteoblasts and promotes the differentiation into osteoclasts in a dose - dependent manner. Helps to understand the causes of bone loss in TZDs in patients with type 2 diabetes.